Protease analysis by neoepitope approach reveals the activation of MMP-9 is achieved proteolytically in a test tissue cartilage model involved in bone formation

ER Lee, L Lamplugh, B Kluczyk… - … of Histochemistry & …, 2006 - journals.sagepub.com
ER Lee, L Lamplugh, B Kluczyk, JS Mort, CP Leblond
Journal of Histochemistry & Cytochemistry, 2006journals.sagepub.com
A principle of regulation of matrix metalloproteinase (MMP) activity has been introduced as
the cysteine-switch mechanism of activation. According to this mechanism, a critical Cys
residue found in the auto-inhibitory propeptide domain of latent proenzyme is important to
determine whether or not activation is turned on or off. The mechanism further allows for
multiple modes of activation. To determine whether or not activation is accomplished
proteolytically within a rat test cartilage model, protease analysis by the neoepitope …
A principle of regulation of matrix metalloproteinase (MMP) activity has been introduced as the cysteine-switch mechanism of activation . According to this mechanism, a critical Cys residue found in the auto-inhibitory propeptide domain of latent proenzyme is important to determine whether or not activation is turned on or off. The mechanism further allows for multiple modes of activation. To determine whether or not activation is accomplished proteolytically within a rat test cartilage model, protease analysis by the neoepitope approach, which relies upon a set of antibodies, was applied. One is used to identify the MMP-9 proenzyme bearing the critical cysteine residue, the other to identify any enzyme present bearing a new NH2-terminus 89FQTFD. This is indicative of MMP-9 lacking the cysteine switch. The antibody set has been applied to frozen tissue sections and analyzed by light and electron microscopic methods. Results reveal that activation of the MMP-9 protease involves limited proteolysis resulting in propeptide domain release. Here we report the observed changes of protease form to indigenous cells and extracellular matrix, thereby making it possible to uncover the features of MMP-9 activation within a specified set of tissue circumstances where a cartilage model is transformed into definitive bone. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials.
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